Korean Journal of Clinical Laboratory Science (대한임상검사과학회지)

Korean Society for Clinical Laboratory Science (대한임상검사과학회)
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Inactivation of Candida albicans Biofilm by Radachlorin-Mediated Photodynamic Therapy

라다클로린으로 매개된 광역학치료에 의한 백색 캔디다 바이오필름의 비활성

  • Kwon, Pil Seung (Department of Clinical Laboratory Science, Wonkwang Health Science University)
  • 권필승 (원광보건대학교 임상병리과)
  • Received : 2015.10.19
  • Accepted : 2015.11.04
  • Published : 2015.12.30
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Abstract

The purpose of this study was to evaluate the in-vitro efficacy of PDT using red light emitting diode (LED) with Radachlorin for biofilm inhibition of clinical Candida albicans isolates. The suspensions containing C. albicans at $9{\times}10^8CFU/mL$ were prepared on yeast nitrogen base containing 5% glucose. The biofilm formation was grown for 3 h after seeding suspensions each 100 ul on a 96-well plate and then supernatant was discarded. Each well was treated with $0.39{\mu}g/mL$ from $50{\mu}g/mL$ concentrations of Radachlorin on adherent biofilm. After a 30-minute incubation, light was irradiated for 30, 60, or 90 minutes using the following light source of wavelength 630 nm LED, at energy densities of 14, 29, and $43J/cm^2$. Afterwards, all supernatant was removed and dried. Adherent cells were stained with safranin O and dried. The cell viability was measured using a microplate reader at 490 nm. Also, a fluorescent signal on C. albicans was observed by saturation of a photosensitizer. In conclusion, a significant inhibition of 72.5% was observed to C. albicans on biofilm at the Radachlorin dose of $50{\mu}g/mL$ with 630 nm LED. The Photosensitizer (Radachlorin) was adequate at 30 minuttes for C. albicans. Overall, the results showed that inhibition of biofilm formation was Radachlorine dose-dependent. The results suggest that PDT, using Radachlorin with 630 nm LED, is able to decrease biofilm formation of C. albicans.

이 연구의 목적은 임상 Candida albicans의 바이오필름억제을 위한 Rhadachlorin과 적색발광다이오드를 이용한 광역학치료의 체외적 효과를 평가하고자 하였다. Candida albicans 부유액을 $9{\times}10^8CFU/mL$을 5% 당이 함유된 YNB배지에 준비하였다. Candida albicans의 바이오필름은 96웰 플레이트에 100 uL씩 분주후 3시간 배양하고 상층액을 제거하였다. 각 웰에 $50{\mu}g/mL$ 부터 $0.39{\mu}g/mL$ 농도로 Rhadachlorin을 부착된 세포위에 처리하였다. 30분간 배양 후 빛을 30, 60, 90분을 630 nm 광원의 LED를 이용하여 조사하였고, 이때 각각의 에너지밀도는 14, 29, $43J/cm^2$이였다. 모든 상층액을 제거후에 건조시켰다. 부착된 세포를 safranin O로 염색하였다. 490 nm 파장으로 마이크로플레이트 리더로 흡광도를 측정하였다. 또한, 광감작제포화에 의해 Candida albicans의 형광시그널을 관찰하였다. 결론적으로 확연한 억제는 Rhadachlorin의 농도 $50{\mu}g/mL$과 630 nm LED치료에서 Candida albicans의 바이오필름의 72.5%가 감소되었다. 또한 광감작제 Rhadachlorin은 Candida albicans의 30분에서 충분한 축적을 보였다. 전체적으로 이 결과들은 바이오필름형성의 억제는 Rhadachlorin의 농도에 의존되었다. 이 결과들은 Rhadachlorin과 630 nm LED의 광역학치료는 Candida albicans의 바이오필름 억제가 가능한 것으로 제의된다.

Keywords

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